Bottom Line:
Fenugreek is a medicinal plant whose seeds are widely used in traditional medicine, mainly for its laxative, galactagogue and antidiabetic effects.The present study was conducted to evaluate the effects of prenatal treatment of fenugreek seeds on the development of sensorimotor functions from birth to young adults.On the other hand, the cross-correlation coefficient in control mice was significantly more negative than in treated animals indicating that alternating patterns are deteriorated in FSAE-treated animals.

ABSTRACTFenugreek is a medicinal plant whose seeds are widely used in traditional medicine, mainly for its laxative, galactagogue and antidiabetic effects. However, consumption of fenugreek seeds during pregnancy has been associated with a range of congenital malformations, including hydrocephalus, anencephaly and spina bifida in humans. The present study was conducted to evaluate the effects of prenatal treatment of fenugreek seeds on the development of sensorimotor functions from birth to young adults. Pregnant mice were treated by gavage with 1 g/kg/day of lyophilized fenugreek seeds aqueous extract (FSAE) or distilled water during the gestational period. Behavioral tests revealed in prenatally treated mice a significant delay in righting, cliff avoidance, negative geotaxis responses and the swimming development. In addition, extracellular recording of motor output in spinal cord isolated from neonatal mice showed that the frequency of spontaneous activity and fictive locomotion was reduced in FSAE-exposed mice. On the other hand, the cross-correlation coefficient in control mice was significantly more negative than in treated animals indicating that alternating patterns are deteriorated in FSAE-treated animals. At advanced age, prenatally treated mice displayed altered locomotor coordination in the rotarod test and also changes in static and dynamic parameters assessed by the CatWalk automated gait analysis system. We conclude that FSAE impairs sensorimotor and coordination functions not only in neonates but also in adult mice. Moreover, spinal neuronal networks are less excitable in prenatally FSAE-exposed mice suggesting that modifications within the central nervous system are responsible, at least in part, for the motor impairments.

pone-0080013-g002: Fenugreek exposure affected swimming performances.We scored the direction of swimming in A. (0; floating, 1; circling, 2; swimming straight or nearly straight, 3), the angle of the head in B (head submerged, 0; nose at the surface, 1; nose and top of head at or above the surface but ears still below the surface, 2; ears half way above the surface, 3; ears completely above the surface,4) and the use of paws in C (0; paddling with all four limbs, 1; paddling with only the hind limbs and the forelimbs remaining motionless, 2). Control mice had a higher performance compared with treated mice, (n = 6). *P < 0.05, **P < 0.01.

Mentions:
We tested the influence of FSAE exposure on sensorimotor tests performed during the postnatal period to evaluate the proper development of the sensorimotor capacities. First, we studied the surface righting reflex that requires the integrity of muscular and motor functions as well as labyrinthine ones. The time required for the surface righting response significantly decreased with age in controls (Spearman Test, P = 0.0049) whereas it did not significantly change in FSAE treated mice (Spearman Test, P = 0.15). Moreover, the righting response was significantly longer in FSAE exposed pups compared with controls at P7 (23.3 ± 3.1 s vs 9.4 ± 3.4 s, (t = 3.7; P< 0.01) and P9 (12.4 ± 3.7 s vs 1.7 ± 0.3 s, t = 2.9, P < 0.05; Figure 1A). Second, we evaluated the geotaxis reflex. When placed on a 35° angle slope, most control mice turned around to orient their body with the head upward within 4s (Figure 1B). This negative geotaxis response was much slower in treated mice as reflected by a ~300% increase in time at both ages tested [P10 (t = 3.9; P < 0.01) and P12 (t = 3.1; P < 0.05)]. Third, we performed the cliff avoidance test: the time taken by treated animals to turn and crawl away from the cliff drop was significantly increased compared with controls (20.8 ± 2.9 s vs 11.9 ± 1.2 s in control; P < 0.01; Figure 1C). We also analyzed the swimming development, with regard to the swimming direction, the treated group had a lower score than the controls at P8 (t = 9; P <0.001), P10 (t= 8.6; P < 0.001) and P12 (t= 9.3; P <0.001) (Figure 2A). There were also significant differences for the swimming angle [P8 (t = 2.9; P <0.05); P10 (t= 4.1; P <0.01)]. However, at P8 and P12, treated mice used all four limbs as frequently as controls. Then at P12, the score became significantly lower in treated animals (t = 23.8; P <0.001) (Figure 2C).

pone-0080013-g002: Fenugreek exposure affected swimming performances.We scored the direction of swimming in A. (0; floating, 1; circling, 2; swimming straight or nearly straight, 3), the angle of the head in B (head submerged, 0; nose at the surface, 1; nose and top of head at or above the surface but ears still below the surface, 2; ears half way above the surface, 3; ears completely above the surface,4) and the use of paws in C (0; paddling with all four limbs, 1; paddling with only the hind limbs and the forelimbs remaining motionless, 2). Control mice had a higher performance compared with treated mice, (n = 6). *P < 0.05, **P < 0.01.

Mentions:
We tested the influence of FSAE exposure on sensorimotor tests performed during the postnatal period to evaluate the proper development of the sensorimotor capacities. First, we studied the surface righting reflex that requires the integrity of muscular and motor functions as well as labyrinthine ones. The time required for the surface righting response significantly decreased with age in controls (Spearman Test, P = 0.0049) whereas it did not significantly change in FSAE treated mice (Spearman Test, P = 0.15). Moreover, the righting response was significantly longer in FSAE exposed pups compared with controls at P7 (23.3 ± 3.1 s vs 9.4 ± 3.4 s, (t = 3.7; P< 0.01) and P9 (12.4 ± 3.7 s vs 1.7 ± 0.3 s, t = 2.9, P < 0.05; Figure 1A). Second, we evaluated the geotaxis reflex. When placed on a 35° angle slope, most control mice turned around to orient their body with the head upward within 4s (Figure 1B). This negative geotaxis response was much slower in treated mice as reflected by a ~300% increase in time at both ages tested [P10 (t = 3.9; P < 0.01) and P12 (t = 3.1; P < 0.05)]. Third, we performed the cliff avoidance test: the time taken by treated animals to turn and crawl away from the cliff drop was significantly increased compared with controls (20.8 ± 2.9 s vs 11.9 ± 1.2 s in control; P < 0.01; Figure 1C). We also analyzed the swimming development, with regard to the swimming direction, the treated group had a lower score than the controls at P8 (t = 9; P <0.001), P10 (t= 8.6; P < 0.001) and P12 (t= 9.3; P <0.001) (Figure 2A). There were also significant differences for the swimming angle [P8 (t = 2.9; P <0.05); P10 (t= 4.1; P <0.01)]. However, at P8 and P12, treated mice used all four limbs as frequently as controls. Then at P12, the score became significantly lower in treated animals (t = 23.8; P <0.001) (Figure 2C).

Bottom Line:
Fenugreek is a medicinal plant whose seeds are widely used in traditional medicine, mainly for its laxative, galactagogue and antidiabetic effects.The present study was conducted to evaluate the effects of prenatal treatment of fenugreek seeds on the development of sensorimotor functions from birth to young adults.On the other hand, the cross-correlation coefficient in control mice was significantly more negative than in treated animals indicating that alternating patterns are deteriorated in FSAE-treated animals.

ABSTRACTFenugreek is a medicinal plant whose seeds are widely used in traditional medicine, mainly for its laxative, galactagogue and antidiabetic effects. However, consumption of fenugreek seeds during pregnancy has been associated with a range of congenital malformations, including hydrocephalus, anencephaly and spina bifida in humans. The present study was conducted to evaluate the effects of prenatal treatment of fenugreek seeds on the development of sensorimotor functions from birth to young adults. Pregnant mice were treated by gavage with 1 g/kg/day of lyophilized fenugreek seeds aqueous extract (FSAE) or distilled water during the gestational period. Behavioral tests revealed in prenatally treated mice a significant delay in righting, cliff avoidance, negative geotaxis responses and the swimming development. In addition, extracellular recording of motor output in spinal cord isolated from neonatal mice showed that the frequency of spontaneous activity and fictive locomotion was reduced in FSAE-exposed mice. On the other hand, the cross-correlation coefficient in control mice was significantly more negative than in treated animals indicating that alternating patterns are deteriorated in FSAE-treated animals. At advanced age, prenatally treated mice displayed altered locomotor coordination in the rotarod test and also changes in static and dynamic parameters assessed by the CatWalk automated gait analysis system. We conclude that FSAE impairs sensorimotor and coordination functions not only in neonates but also in adult mice. Moreover, spinal neuronal networks are less excitable in prenatally FSAE-exposed mice suggesting that modifications within the central nervous system are responsible, at least in part, for the motor impairments.